APL Bioengineering最新文献

{"title":"Air-liquid intestinal cell culture allows <i>in situ</i> rheological characterization of intestinal mucus.","authors":"Pamela C Cai, Margaret Braunreuther, Audrey Shih, Andrew J Spakowitz, Gerald G Fuller, Sarah C Heilshorn","doi":"10.1063/5.0187974","DOIUrl":"10.1063/5.0187974","url":null,"abstract":"<p><p>Intestinal health heavily depends on establishing a mucus layer within the gut with physical properties that strike a balance between being sufficiently elastic to keep out harmful pathogens yet viscous enough to flow and turnover the contents being digested. Studies investigating dysfunction of the mucus layer in the intestines are largely confined to animal models, which require invasive procedures to collect the mucus fluid. In this work, we develop a nondestructive method to study intestinal mucus. We use an air-liquid interface culture of primary human intestinal epithelial cells that exposes their apical surface to allow <i>in situ</i> analysis of the mucus layer. Mucus collection is not only invasive but also disrupts the mucus microstructure, which plays a crucial role in the interaction between mucus and the gut microbiome. Therefore, we leverage a noninvasive rheology technique that probes the mechanical properties of the mucus without removal from the culture. Finally, to demonstrate biomedical uses for this cell culture system, we characterize the biochemical and biophysical properties of intestinal mucus due to addition of the cytokine IL-13 to recapitulate the gut environment of <i>Nippostrongylus brasiliensis</i> infection.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 2","pages":"026112"},"PeriodicalIF":6.6,"publicationDate":"2024-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11078553/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140892808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nondestructive, quantitative viability analysis of 3D tissue cultures using machine learning image segmentation.","authors":"Kylie J Trettner, Jeremy Hsieh, Weikun Xiao, Jerry S H Lee, Andrea M Armani","doi":"10.1063/5.0189222","DOIUrl":"10.1063/5.0189222","url":null,"abstract":"<p><p>Ascertaining the collective viability of cells in different cell culture conditions has typically relied on averaging colorimetric indicators and is often reported out in simple binary readouts. Recent research has combined viability assessment techniques with image-based deep-learning models to automate the characterization of cellular properties. However, further development of viability measurements to assess the continuity of possible cellular states and responses to perturbation across cell culture conditions is needed. In this work, we demonstrate an image processing algorithm for quantifying features associated with cellular viability in 3D cultures without the need for assay-based indicators. We show that our algorithm performs similarly to a pair of human experts in whole-well images over a range of days and culture matrix compositions. To demonstrate potential utility, we perform a longitudinal study investigating the impact of a known therapeutic on pancreatic cancer spheroids. Using images taken with a high content imaging system, the algorithm successfully tracks viability at the individual spheroid and whole-well level. The method we propose reduces analysis time by 97% in comparison with the experts. Because the method is independent of the microscope or imaging system used, this approach lays the foundation for accelerating progress in and for improving the robustness and reproducibility of 3D culture analysis across biological and clinical research.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"016121"},"PeriodicalIF":6.6,"publicationDate":"2024-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10985731/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140872559","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Erratum: Publisher's Note: \"Magneto-responsive hyaluronan hydrogel for hyperthermia and bioprinting: Magnetic, rheological properties and biocompatibility\" [APL Bioeng. <b>7</b>, 036113 (2023)].","authors":"L Vítková, N Kazantseva, L Musilová, P Smolka, K Valášková, K Kocourková, M Humeník, A Minařík, P Humpolíček, A Mráček, I Smolková","doi":"10.1063/5.0207818","DOIUrl":"https://doi.org/10.1063/5.0207818","url":null,"abstract":"<p><p>[This corrects the article DOI: 10.1063/5.0147181.].</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"019901"},"PeriodicalIF":6.0,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10954346/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140177067","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Differential roles of normal and lung cancer-associated fibroblasts in microvascular network formation.","authors":"Naveen R Natesh, Pankaj Mogha, Alan Chen, Scott J Antonia, Shyni Varghese","doi":"10.1063/5.0188238","DOIUrl":"10.1063/5.0188238","url":null,"abstract":"<p><p>Perfusable microvascular networks offer promising three-dimensional <i>in vitro</i> models to study normal and compromised vascular tissues as well as phenomena such as cancer cell metastasis. Engineering of these microvascular networks generally involves the use of endothelial cells stabilized by fibroblasts to generate robust and stable vasculature. However, fibroblasts are highly heterogenous and may contribute variably to the microvascular structure. Here, we study the effect of normal and cancer-associated lung fibroblasts on the formation and function of perfusable microvascular networks. We examine the influence of cancer-associated fibroblasts on microvascular networks when cultured in direct (juxtacrine) and indirect (paracrine) contacts with endothelial cells, discovering a generative inhibition of microvasculature in juxtacrine co-cultures and a functional inhibition in paracrine co-cultures. Furthermore, we probed the secreted factors differential between cancer-associated fibroblasts and normal human lung fibroblasts, identifying several cytokines putatively influencing the resulting microvasculature morphology and functionality. These findings suggest the potential contribution of cancer-associated fibroblasts in aberrant microvasculature associated with tumors and the plausible application of such <i>in vitro</i> platforms in identifying new therapeutic targets and/or agents that can prevent formation of aberrant vascular structures.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"016120"},"PeriodicalIF":6.0,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10959556/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140207857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Asymmetrical positioning of cell organelles reflects the cell chirality of mouse myoblast cells.","authors":"Zeina Hachem, Courtney Hadrian, Lina Aldbaisi, Muslim Alkaabi, Leo Q Wan, Jie Fan","doi":"10.1063/5.0189401","DOIUrl":"10.1063/5.0189401","url":null,"abstract":"<p><p>Cell chirality is crucial for the chiral morphogenesis of biological tissues, yet its underlying mechanism remains unclear. Cell organelle polarization along multiple axes in a cell body, namely, apical-basal, front-rear, and left-right, is known to direct cell behavior such as orientation, rotation, and migration. Among these axes, the left-right bias holds significant sway in determining the chiral directionality of these behaviors. Normally, mouse myoblast (C2C12) cells exhibit a strong counterclockwise chirality on a ring-shaped micropattern, whereas they display a clockwise dominant chirality under Latrunculin A treatment. To investigate the relationship between multicellular chirality and organelle positioning in single cells, we studied the left-right positioning of cell organelles under distinct cell chirality in single cells via micropatterning technique, fluorescent microscopy, and imaging analysis. We found that on a \"T\"-shaped micropattern, a C2C12 cell adopts a triangular shape, with its nucleus-centrosome axis pointing toward the top-right direction of the \"T.\" Several other organelles, including the Golgi apparatus, lysosomes, actin filaments, and microtubules, showed a preference to polarize on one side of the axis, indicating the universality of the left-right asymmetrical organelle positioning. Interestingly, upon reversing cell chirality with Latrunculin A, the organelles correspondingly reversed their left-right positioning bias, as suggested by the consistently biased metabolism and contractile properties at the leading edge. This left-right asymmetry in organelle positioning may help predict cell migration direction and serve as a potential marker for identifying cell chirality in biological models.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"016119"},"PeriodicalIF":6.0,"publicationDate":"2024-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10942803/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140144278","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Static magnetic fields in regenerative medicine.","authors":"Wenjing Xie, Chao Song, Ruowen Guo, Xin Zhang","doi":"10.1063/5.0191803","DOIUrl":"10.1063/5.0191803","url":null,"abstract":"<p><p>All organisms on Earth live in the weak but ubiquitous geomagnetic field. Human beings are also exposed to magnetic fields generated by multiple sources, ranging from permanent magnets to magnetic resonance imaging (MRI) in hospitals. It has been shown that different magnetic fields can generate various effects on different tissues and cells. Among them, stem cells appear to be one of the most sensitive cell types to magnetic fields, which are the fundamental units of regenerative therapies. In this review, we focus on the bioeffects of static magnetic fields (SMFs), which are related to regenerative medicine. Most reports in the literature focus on the influence of SMF on bone regeneration, wound healing, and stem cell production. Multiple aspects of the cellular events, including gene expression, cell signaling pathways, reactive oxygen species, inflammation, and cytoskeleton, have been shown to be affected by SMFs. Although no consensus yet, current evidence indicates that moderate and high SMFs could serve as a promising physical tool to promote bone regeneration, wound healing, neural differentiation, and dental regeneration. All <i>in vivo</i> studies of SMFs on bone regeneration and wound healing have shown beneficial effects, which unravel the great potential of SMFs in these aspects. More mechanistic studies, magnetic field parameter optimization, and clinical investigations on human bodies will be imperative for the successful clinical applications of SMFs in regenerative medicine.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"011503"},"PeriodicalIF":6.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10939708/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140132792","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Terahertz <i>in vivo</i> imaging of human skin: Toward detection of abnormal skin pathologies.","authors":"X Qi, K Bertling, J Torniainen, F Kong, T Gillespie, C Primiero, M S Stark, P Dean, D Indjin, L H Li, E H Linfield, A G Davies, M Brünig, T Mills, C Rosendahl, H P Soyer, A D Rakić","doi":"10.1063/5.0190573","DOIUrl":"10.1063/5.0190573","url":null,"abstract":"<p><p>Terahertz (THz) imaging has long held promise for skin cancer detection but has been hampered by the lack of practical technological implementation. In this article, we introduce a technique for discriminating several skin pathologies using a coherent THz confocal system based on a THz quantum cascade laser. High resolution <i>in vivo</i> THz images (with diffraction limited to the order of 100 <i>μ</i>m) of several different lesion types were acquired and compared against one another using the amplitude and phase values. Our system successfully separated pathologies using a combination of phase and amplitude information and their respective surface textures. The large scan field (50 × 40 mm) of the system allows macroscopic visualization of several skin lesions in a single frame. Utilizing THz imaging for dermatological assessment of skin lesions offers substantial additional diagnostic value for clinicians. THz images contain information complementary to the information contained in the conventional digital images.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"016117"},"PeriodicalIF":6.0,"publicationDate":"2024-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10932572/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140111833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineered tissue geometry and Plakophilin-2 regulate electrophysiology of human iPSC-derived cardiomyocytes.","authors":"Daniel W Simmons, Ganesh Malayath, David R Schuftan, Jingxuan Guo, Kasoorelope Oguntuyo, Ghiska Ramahdita, Yuwen Sun, Samuel D Jordan, Mary K Munsell, Brennan Kandalaft, Missy Pear, Stacey L Rentschler, Nathaniel Huebsch","doi":"10.1063/5.0160677","DOIUrl":"10.1063/5.0160677","url":null,"abstract":"<p><p>Engineered heart tissues have been created to study cardiac biology and disease in a setting that more closely mimics <i>in vivo</i> heart muscle than 2D monolayer culture. Previously published studies suggest that geometrically anisotropic micro-environments are crucial for inducing \"<i>in vivo</i> like\" physiology from immature cardiomyocytes. We hypothesized that the degree of cardiomyocyte alignment and prestress within engineered tissues is regulated by tissue geometry and, subsequently, drives electrophysiological development. Thus, we studied the effects of tissue geometry on electrophysiology of micro-heart muscle arrays (<i>μ</i>HM) engineered from human induced pluripotent stem cells (iPSCs). Elongated tissue geometries elicited cardiomyocyte shape and electrophysiology changes led to adaptations that yielded increased calcium intake during each contraction cycle. Strikingly, pharmacologic studies revealed that a threshold of prestress and/or cellular alignment is required for sodium channel function, whereas L-type calcium and rapidly rectifying potassium channels were largely insensitive to these changes. Concurrently, tissue elongation upregulated sodium channel (Na_V1.5) and gap junction (Connexin 43, Cx43) protein expression. Based on these observations, we leveraged elongated <i>μ</i>HM to study the impact of loss-of-function mutation in Plakophilin 2 (PKP2), a desmosome protein implicated in arrhythmogenic disease. Within <i>μ</i>HM, PKP2 knockout cardiomyocytes had cellular morphology similar to what was observed in isogenic controls. However, PKP2^-/- tissues exhibited lower conduction velocity and no functional sodium current. PKP2 knockout <i>μ</i>HM exhibited geometrically linked upregulation of sodium channel but not Cx43, suggesting that post-translational mechanisms, including a lack of ion channel-gap junction communication, may underlie the lower conduction velocity observed in tissues harboring this genetic defect. Altogether, these observations demonstrate that simple, scalable micro-tissue systems can provide the physiologic stresses necessary to induce electrical remodeling of iPS-CM to enable studies on the electrophysiologic consequences of disease-associated genomic variants.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"016118"},"PeriodicalIF":6.6,"publicationDate":"2024-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10932571/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140111832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advances in cancer mechanobiology: Metastasis, mechanics, and materials.","authors":"Abigail J Clevenger, Maygan K McFarlin, John Paul M Gorley, Spencer C Solberg, Anirudh K Madyastha, Shreya A Raghavan","doi":"10.1063/5.0186042","DOIUrl":"10.1063/5.0186042","url":null,"abstract":"<p><p>Within the tumor microenvironment (TME), tumor cells are exposed to numerous mechanical forces, both internally and externally, which contribute to the metastatic cascade. From the initial growth of the tumor to traveling through the vasculature and to the eventual colonization of distant organs, tumor cells are continuously interacting with their surroundings through physical contact and mechanical force application. The mechanical forces found in the TME can be simplified into three main categories: (i) shear stress, (ii) tension and strain, and (iii) solid stress and compression. Each force type can independently impact tumor growth and progression. Here, we review recent bioengineering strategies, which have been employed to establish the connection between mechanical forces and tumor progression. While many cancers are explored in this review, we place great emphasis on cancers that are understudied in their response to mechanical forces, such as ovarian and colorectal cancers. We discuss the major steps of metastatic transformation and present novel, recent advances in model systems used to study how mechanical forces impact the study of the metastatic cascade. We end by summarizing systems that incorporate multiple forces to expand the complexity of our understanding of how tumor cells sense and respond to mechanical forces in their environment. Future studies would also benefit from the inclusion of time or the aspect of mechanical memory to further enhance this field. While the knowledge of mechanical forces and tumor metastasis grows, developing novel materials and <i>in vitro</i> systems are essential to providing new insight into predicting, treating, and preventing cancer progression and metastasis.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"011502"},"PeriodicalIF":6.0,"publicationDate":"2024-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10917464/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140050588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanoregulation of MSC spheroid immunomodulation.","authors":"Victoria L Thai, Sabrina Mierswa, Katherine H Griffin, Joel D Boerckel, J Kent Leach","doi":"10.1063/5.0184431","DOIUrl":"10.1063/5.0184431","url":null,"abstract":"<p><p>Mesenchymal stromal cells (MSCs) are widely used in cell-based therapies and tissue regeneration for their potent secretome, which promotes host cell recruitment and modulates inflammation. Compared to monodisperse cells, MSC spheroids exhibit improved viability and increased secretion of immunomodulatory cytokines. While mechanical stimulation of monodisperse cells can increase cytokine production, the influence of mechanical loading on MSC spheroids is unknown. Here, we evaluated the effect of controlled, uniaxial cyclic compression on the secretion of immunomodulatory cytokines by human MSC spheroids and tested the influence of load-induced gene expression on MSC mechanoresponsiveness. We exposed MSC spheroids, entrapped in alginate hydrogels, to three cyclic compressive regimes with varying stress (L) magnitudes (i.e., 5 and 10 kPa) and hold (H) durations (i.e., 30 and 250 s) L5H30, L10H30, and L10H250. We observed changes in cytokine and chemokine expression dependent on the loading regime, where higher stress regimes tended to result in more exaggerated changes. However, only MSC spheroids exposed to L10H30 induced human THP-1 macrophage polarization toward an M2 phenotype compared to static conditions. Static and L10H30 loading facilitated a strong, interlinked F-actin arrangement, while L5H30 and L10H250 disrupted the structure of actin filaments. This was further examined when the actin cytoskeleton was disrupted via Y-27632. We observed downregulation of YAP-related genes, and the levels of secreted inflammatory cytokines were globally decreased. These findings emphasize the essential role of mechanosignaling in mediating the immunomodulatory potential of MSC spheroids.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":"8 1","pages":"016116"},"PeriodicalIF":6.0,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10908560/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140022891","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}